Effect of miR-21 in mesenchymal stem cells-derived extracellular vesicles behavior.

BACKGROUND: A challenging new branch of research related to aging-associated diseases is the identification of miRNAs capable of modulating the senescence-associated secretory phenotype (SASP) which characterizes senescent cells and contributes to driving inflammation. METHODS: Mesenchymal stem cells (MSC) from human umbilical cord stroma were stable modified using lentivirus transduction to inhibit miR-21-5p and shotgun proteomic analysis was performed in the MSC-derived extracellular vesicles (EV) to check the effect of miR-21 inhibition in their protein cargo. Besides, we studied the paracrine effect of those modified extracellular vesicles and also their effect on SASP. RESULTS: Syndecan-1 (SDC1) was the most decreased protein in MSC-miR21--derived EV, and it was involved in inflammation and EV production. MSC-miR21--derived EV were found to produce a statistically significant inhibitory effect on SASP and inflammaging markers expression in receptor cells, and in the opposite way, these receptor cells increased their SASP and inflammaging expression statistically significantly when treated with MSC-miR-21+-derived EV. CONCLUSION: This work demonstrates the importance of miR-21 in inflammaging and its role in SASP through SDC1.

Study of Ferroptosis Transmission by Small Extracellular Vesicles in Epithelial Ovarian Cancer Cells.

Epithelial ovarian cancer (EOC) is the most lethal gynecological cancer. The current treatment for EOC involves surgical debulking of the tumors followed by a combination of chemotherapy. While most patients achieve complete remission, many EOCs will recur and develop chemo-resistance. The cancer cells can adapt to several stress stimuli, becoming resistant. Because of this, new ways to fight resistant cells during the disease are being studied. However, the clinical outcomes remain unsatisfactory. Recently, ferroptosis, a novel form of regulated cell death trigged by the accumulation of iron and toxic species of lipid metabolism in cells, has emerged as a promising anti-tumor strategy for EOC treatment. This process has a high potential to become a complementary treatment to the current anti-tumor strategies to eliminate resistant cells and to avoid relapse. Cancer cells, like other cells in the body, release small extracellular vesicles (sEV) that allow the transport of substances from the cells themselves to communicate with their environment. To achieve this, we analyzed the capacity of epithelial ovarian cancer cells (OVCA), treated with ferroptosis inducers, to generate sEV, assessing their size and number, and study the transmission of ferroptosis by sEV. Our results reveal that OVCA cells treated with ferroptotic inducers can modify intercellular communication by sEV, inducing cell death in recipient cells. Furthermore, these receptor cells are able to generate a greater amount of sEV, contributing to a much higher ferroptosis paracrine transmission. Thus, we discovered the importance of the sEV in the communication between cells in OVCA, focusing on the ferroptosis process. These findings could be the beginning form to study the molecular mechanism ferroptosis transmission through sEV.

Therapy free of cells vs human mesenchymal stem cells from umbilical cord stroma to treat the inflammation in OA.

Osteoarthritis (OA) is closely linked to the increase in the number of senescent cells in joint tissues, and the senescence-associated secretory phenotype (SASP) is implicated in cartilage degradation. In the last decade, extracellular vesicles (EV) in combination with the use of miRNAs to modify post-transcriptional expressions of multiple genes have shown their utility in new therapies to treat inflammatory diseases. This work delves into the anti-inflammatory effect of extracellular vesicles derived from mesenchymal stem cells (MSC) previously modified to inhibit the expression of miR-21. We compare the efficacy of two treatments, MSC with their miR-21 inhibited through lentiviral transfection and their EV, against inflammation in a new OA animal model. The modified MSC and their EV were intraperitoneally injected in an OA animal model twice. One month after treatment, we checked which therapy was the most effective to reduce inflammation compared with animals untreated. Treated OA model sera were analyzed for cytokines and chemokines. Subsequently, different organs were analyzed to validate the results obtained. EV were the most effective treatment to reduce chemokines and cytokines in serum of OA animals as well as SASP, in their organs checked by proteomic and genomic techniques, compared with MSC alone in a statistically significant way. In conclusion, MSC-miR-21--derived EV showed a higher therapeutic potential in comparison with MSCs-miR-21-. They ameliorate the systemic inflammation through inactivation of ERK1/2 pathway in OA in vivo model. Workflow of the realization of the animal model of OA by injecting cells into the joint cavity of the left knee of the animals, which produces an increase in serum cytokines and chemokines in the animals in addition to the increase in SASP and markers of inflammation. Inhibition of miR-21 in MSCs, from the stroma of the human umbilical cord, by lentivirus and extraction of their EVs by ultracentrifugation. Finally, application of MSC therapy with its miR-21 inhibited or its EVs produces a decrease in serum cytokines and chemokines in the treated animals, in addition to an increase in SASP and markers of inflammation. The cell-free therapy being the one that produces a greater decrease in the parameters studied.

Action Mechanisms of Small Extracellular Vesicles in Inflammaging.

The accumulation process of proinflammatory components in the body due to aging influences intercellular communication and is known as inflammaging. This biological mechanism relates the development of inflammation to the aging process. Recently, it has been reported that small extracellular vesicles (sEVs) are mediators in the transmission of paracrine senescence involved in inflammatory aging. For this reason, their components, as well as mechanisms of action of sEVs, are relevant to develop a new therapy called senodrugs (senolytics and senomorphic) that regulates the intercellular communication of inflammaging. In this review, we include the most recent and relevant studies on the role of sEVs in the inflammatory aging process and in age-related diseases such as cancer and type 2 diabetes.

Evolución de los pacientes con enfermedad hepática crítica / Prognosis of patients with critical hepatic disease

INTRODUCTION: The acute liver failure on chronic (ACLF), is an entity, whose recognition is increasing. The ACLF and CLIF OF indexes have been recently presented with the objective of predicting mortality in this kind of patients. MATERIAL AND METHODS: All patients admitted to the Ramón y Cajal University Hospital diagnosed of acute liver failure on chronic during 2016 and 2017 were collected. We collect the scores: SOFA, CLIF, APACHE II, SAPS II and ACLF score in patients admitted to the ICU by comparing them with each other and define which stages have worse prognosis. RESULTS: A total of 46 patients were collected. The study presents an intra ICU mortality of 31% (15/46) and a six-month mortality of 59.6% (28/46). Patients classified as death, present ACLF values ​​at admission (49.5 vs 60 p = 0.001), and at three days (46.66 vs 59.4 p = 0.001) higher than survivors. In the analysis of the ROC curve, the area under the curve in relation to six-month mortality is higher in the ACLF index (0.8) compared to the MELD (0.69) SOFA (0.66) SAPS II (0.69) or APACHE II (0.65). Patients with ACLF indexes above 65 had an intra UCI mortality of 54%, however, mortality at 6 months is 90%. Patients with ACLF values ​​greater than 65 present mean values ​​of lactic acid, leukocytes, INR or bilirubin higher than those under 65 in a statistically significant manner. CONCLUSIONS: The data presented in this study suggest that the ACLF index works as an adequate predictor of intra-ICU mortality and at 6 months.

INTRODUCCIÓN: El fallo hepático agudo sobre crónico es una entidad cuyo reconocimiento va en aumento. Los índices ACLF y CLIF OF, han sido presentados recientemente con el objetivo de predecir la mortalidad en este tipo de enfermos. MATERIAL Y MÉTODOS: Se recogen todos los pacientes ingresados en una unidad de cuidados intensivos (UCI) de un hospital terciario universitario, diagnosticados de fallo hepático agudo sobre crónico durante 2016 y 2017. Recogemos los índices SOFA, CLIF, APACHE II, SAPS II Y ACLF en pacientes ingresados en UCI comparándolos entre sí. Definimos que estadios presentan peor pronóstico. RESULTADOS: Se analizan un total de 46 pacientes. El estudio presenta una mortalidad intra-UCI del 31% (15/46) y una mortalidad a los seis meses de 59,6% (28/46). Los pacientes clasificados como éxitus presentan valores ACLF al ingreso (49,5 vs 60 p = 0,001), a los tres días (46,66 vs 59,4 p = 0,001) superiores a los supervivientes. En el análisis de la curva COR, el área bajo la curva en relación a la mortalidad a los seis meses, es superior en el índice ACLF (0,8) en comparación con el MELD (0,69) SOFA (0,66) SAPS II (0,69) o APACHE II (0,65). Los pacientes con índices ACLF superiores a 65 presentaban una mortalidad intra-UCI del 54% sin embargo, la mortalidad a los 6 meses es del 90%. Los pacientes con valores ACLF superiores a 65 presentan a su vez valores medios de láctico, leucocitos, INR o bilirrubina mayores de forma estadísticamente significativa. CONCLUSIONES: Los datos presentados en este estudio sugieren que el índice ACLF funciona como un adecuado predictor de mortalidad intra-UCI y a los 6 meses.

Análisis función ventricular derecha, función renal y fluidos tras resucitación en pacientes con ventilación mecánica / Right ventricular, renal function, and fluids balance analysis after resuscitation in patients on mechanical ventilation

INTRODUCTION: Hemodynamic optimization is a main goal in the management of critically ill patients. Right ventricular function, renal failure and fluid balance are part of this process. Our goal is to analysis those, after the initial resuscitation. MATERIAL AND METHODS: A prospective, observational study was performed with all the patients admitted to a Medical Service of Intensive Care of a Tertiary, University Hospital. All patients analyzed required mechanical ventilation as a support for their underling pathology. All consecutive patients admitted to the unit under mechanical ventilation were collected, in the absence of shock, with the expectation of remaining under mechanical ventilation for at least 24 more hours after data collection. The incidence of right ventricular failure according to the defined parameters, renal failure, and fluid balance were described. We had described its association with mechanical ventilation and mortality. RESULTS: A total of 30 patients were selected. Right ventricular failure was observed in 16.6% of patients (5/30). There was no statistically significant association with the need of tracheotomy, renal failure or mortality. It was not associated with longer average stay, days of mechanical ventilation or higher severity scores. 40% of the patients presented acute renal failure. Renal failure was not associated statistically significant with the need of tracheotomy or failure in scheduled extubation, however, it was associated with higher mortality. Patients requiring mechanical ventilation with normal creatinine values ​​after initial resuscitation, had a mortality rate of 28.5% in comparison to patients who had altered values in which the mortality rate was 77.7%, (p < 0.018). Regarding the post-resuscitation net fluid balance, no statistically significant differences were found in the comparison of means between survivors and non survivors, with renal failure or right ventricular failure. No even was it associated with higher mortality. CONCLUSIONS: Right ventricular failure, despite not being associated with mortality, days of mechanical ventilation or failure in extubation in a statistically significant way, presents an incidence of 16.6% in patients connected to mechanical ventilation admitted to a polyvalent icu. Acute renal failure is associated in a statistically significant way with mortality in our sample. We had not found association between fluid overload and renal failure in our sample.

INTRODUCCIÓN: La optimización hemodinámica es piedra angular en el manejo del enfermo crítico. La función ventricular derecha, el fallo renal y el balance hídrico son parte de este proceso. Nuestro objetivo es su análisis tras la resucitación inicial. MATERIAL Y MÉTODOS: Se realiza un estudio prospectivo, observacional, con todos los enfermos que ingresan en un Servicio médico de Medicina Intensiva de un Hospital Terciario y Universitario. Todos los enfermos analizados precisan ventilación mecánica como soporte de su patología. Se recogen todos los enfermos consecutivos ingresados en la unidad bajo ventilación mecánica, en ausencia de shock, con previsión de permanecer bajo ventilación mecánica al menos 24 horas más tras la recogida de datos. Se describe la incidencia de fallo ventricular derecho según los parámetros definidos, fallo renal, balance. Describiremos también su asociación con ventilación mecánica, mortalidad. RESULTADOS: Se seleccionaron un total de 30 pacientes. Se objetivó fallo ventricular derecho en el 16,6% de los pacientes (5/30). No se asoció de forma estadísticamente significativa a la necesidad de traqueotomía, fallo renal o mortalidad. Tampoco se asoció a mayor estancia media, días de ventilación mecánica o índices de gravedad mayores. El 40% de los pacientes presentaron fallo renal agudo. El fallo renal no se asocia de forma estadísticamente significativa con la necesidad de traqueotomía ni fracaso en la extubación, sin embargo, se asocia a mayor mortalidad. Los pacientes que precisan ventilación mecánica y presentan valores de creatinina normales tras resucitación inicial, cuentan con una tasa de mortalidad del 28,5% en comparación con aquellos pacientes que presentan cifras alteradas en las cuales la tasa de mortalidad es del 77,7%, (p < 0,018). Con respecto al balance hídrico neto post resucitación, no se encontraron diferencias estadísticamente significativas en la comparación de medias de los supervivientes, con fallo renal o fallo en ventrículo derecho. Tampoco se asoció a mayor mortalidad. CONCLUSIONES: El fallo ventricular derecho, pese a no asociarse a la mortalidad, días de ventilación mecánica o fallo en la extubación de forma estadísticamente significativa, presenta una incidencia del 16,6% en los enfermos conectados a ventilación mecánica ingresados en una uci polivalente. El fallo renal agudo sí se asocia de forma estadísticamente significativa a la mortalidad en nuestra muestra. No hemos encontrado asociación entre sobrecarga hídrica y fallo renal en nuestro estudio.

Multivariate Discretization Based on Evolutionary Cut Points Selection for Classification.

Discretization is one of the most relevant techniques for data preprocessing. The main goal of discretization is to transform numerical attributes into discrete ones to help the experts to understand the data more easily, and it also provides the possibility to use some learning algorithms which require discrete data as input, such as Bayesian or rule learning. We focus our attention on handling multivariate classification problems, where high interactions among multiple attributes exist. In this paper, we propose the use of evolutionary algorithms to select a subset of cut points that defines the best possible discretization scheme of a data set using a wrapper fitness function. We also incorporate a reduction mechanism to successfully manage the multivariate approach on large data sets. Our method has been compared with the best state-of-the-art discretizers on 45 real datasets. The experiments show that our proposed algorithm overcomes the rest of the methods producing competitive discretization schemes in terms of accuracy, for C4.5, Naive Bayes, PART, and PrUning and BuiLding Integrated in Classification classifiers; and obtained far simpler solutions.

Combined approach of hematological biomarkers and plasma protein SAA for improvement of radiation dose assessment triage in biodosimetry applications.

Early treatment of populations exposed to ionizing radiation requires accurate and rapid biodosimetry with a precision as high as possible to determine an individual's exposure level and risk for morbidity and mortality. The purpose of this study was to evaluate the utility of multiple blood biomarkers for early-response assessment of radiation exposure using a murine (BALB/c, males) in vivo radiation model. Present results for mice exposed to whole-body Co gamma-rays (0.1 Gy min) over a broad dose range (0-7 Gy) demonstrate at 24 h after exposure: 1) dose-dependent increase in the acute phase protein serum amyloid A or SAA; 2) dose-dependent changes in blood cell counts (lymphocytes, neutrophils, and ratio of neutrophils to lymphocytes); 3) SAA results coupled with peripheral blood cell counts analyzed with use of multivariate discriminant analysis established very successful separation of irradiated animals; 4) an enhanced separation as the number of biomarkers increased. These results also demonstrate proof-in-concept that plasma protein SAA shows promise as a complimentary approach to conventional biodosimetry for early assessment of radiation exposures and, coupled with peripheral blood cell counts, provides early diagnostic information to effectively manage radiation casualty incidents.